Yanhua Peng1, Michel M Murr. 1. James A. Haley Veterans Affairs Medical Center, Department of Surgery, University of South Florida Health Sciences Center, C/O Tampa General Hospital, Tampa, FL 33601, USA.
Abstract
BACKGROUND: Kupffer cells have been implicated in the pathogenesis of various liver diseases. Primary cultures of Kupffer cells have a very limited life span, tend to de-differentiate and become senescent, and therefore are not suitable for cell signaling studies. AIM: To establish immortalized rat Kupffer cell lines that facilitate mechanistic studies of cell signaling and signal transduction. METHODS: Rat Kupffer cells were sub-cultured with EGF to obtain rat Kupffer Cell line 1 (RKC1), and subsequently transfected with Simian Virus 40 Large T-antigen expression vector to obtain rat Kupffer Cell line 2 (RKC2). RESULTS: RKC1 and RKC2 are similar to primary Kupffer cells as they express the molecular markers ED1, ED2, ED3, and F4/80, and upregulate TNF-alpha, IL-6, IL-1beta, Fas /FasL, and NF-kappaB, as well as TLR4 in response to LPS or pancreatic elastase. Additionally, RKC1 and RKC2 maintain phagocytic properties of latex beads and exhibit increased telomerase and stabilized p53 activity. CONCLUSION: Immortalized RKC1 and RKC2 cells maintain properties of primary Kupffer cells and can be valuable tools in evaluating the role of Kupffer cells in immune diseases and in liver-cell based drug discovery.
BACKGROUND: Kupffer cells have been implicated in the pathogenesis of various liver diseases. Primary cultures of Kupffer cells have a very limited life span, tend to de-differentiate and become senescent, and therefore are not suitable for cell signaling studies. AIM: To establish immortalized rat Kupffer cell lines that facilitate mechanistic studies of cell signaling and signal transduction. METHODS:Rat Kupffer cells were sub-cultured with EGF to obtain rat Kupffer Cell line 1 (RKC1), and subsequently transfected with Simian Virus 40 Large T-antigen expression vector to obtain rat Kupffer Cell line 2 (RKC2). RESULTS: RKC1 and RKC2 are similar to primary Kupffer cells as they express the molecular markers ED1, ED2, ED3, and F4/80, and upregulate TNF-alpha, IL-6, IL-1beta, Fas /FasL, and NF-kappaB, as well as TLR4 in response to LPS or pancreatic elastase. Additionally, RKC1 and RKC2 maintain phagocytic properties of latex beads and exhibit increased telomerase and stabilized p53 activity. CONCLUSION: Immortalized RKC1 and RKC2 cells maintain properties of primary Kupffer cells and can be valuable tools in evaluating the role of Kupffer cells in immune diseases and in liver-cell based drug discovery.
Authors: Zheng Shen; Joanne M Ajmo; Christopher Q Rogers; Xiaomei Liang; Lisa Le; Michel M Murr; Yanhua Peng; Min You Journal: Am J Physiol Gastrointest Liver Physiol Date: 2009-03-19 Impact factor: 4.052
Authors: Yanhua Peng; Drew Rideout; Steven Rakita; Mini Sajan; Robert Farese; Min You; Michel M Murr Journal: J Gastrointest Surg Date: 2009-09-18 Impact factor: 3.452
Authors: Simone Merlin; Kuldeep K Bhargava; Gabriella Ranaldo; Diego Zanolini; Christopher J Palestro; Laura Santambrogio; Maria Prat; Antonia Follenzi; Sanjeev Gupta Journal: Am J Pathol Date: 2016-01-07 Impact factor: 4.307
Authors: Karen Wallace; David E Cowie; Dimitrios K Konstantinou; Stephen J Hill; Torunn E Tjelle; Andrew Axon; Matthew Koruth; Steven A White; Harald Carlsen; Derek A Mann; Matthew C Wright Journal: J Steroid Biochem Mol Biol Date: 2010-04-21 Impact factor: 4.292